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猪呼吸道冠状病毒S1基因的克隆与序列分析
引用本文:邱深本,王磊,苏丹萍,黄爱芳,罗映霞,贺东生.猪呼吸道冠状病毒S1基因的克隆与序列分析[J].中国动物检疫,2009,26(6):36-38.
作者姓名:邱深本  王磊  苏丹萍  黄爱芳  罗映霞  贺东生
作者单位:1. 广东科贸职业学院生物技术系,广州,510430
2. 华南农业大学兽医学院,广东省人兽共患病重点实验室,广州,510642
摘    要:以猪呼吸道冠状病毒(PRCV)AR310毒株为研究对象,观察PRCV在两种不同细胞中的生长繁殖情况,在猪肾原代细胞中生长良好,在PK15细胞中未见生长。参考GenBbank中已发表的基因序列,设计和合成一对引物,通过RT-PCR扩增出PRCVAR310株的S1基因,克隆到pMD18-T载体中并测序。结果表明与PRCVDQ811787核苷酸同源性为100%,与M94096、M94097的同源性分别为95.2%和95.0%;氨基酸序列分析表明,试验测序的PRCVS1基因氨基酸序列与Genebank中DQ811787同源性为100%,与M94096、M94097的同源性较高分别为95.2%和95.6%。

关 键 词:PRCV  S1基因  克隆  序列分析

Clone and Nucleotide Analysis of PRCV D1 Gene
QIU Shenben,WANG Lei,Su Danping,HUANG Aifang,LUO Yingxi,HE Dongsheng.Clone and Nucleotide Analysis of PRCV D1 Gene[J].China Journal Of Animal Quarantine,2009,26(6):36-38.
Authors:QIU Shenben  WANG Lei  Su Danping  HUANG Aifang  LUO Yingxi  HE Dongsheng
Institution:1. Guangdong Vocational College of Science and Trade, Guangzhou 510430 China;2.College of Veterinary Medicine, South China Agricultural University, Guangzhou 510640 China)
Abstract:The propagation of porcine respiratory coronavirus (PRCV) AR310 stain in 2 cell lines was studied. It showed that the primary porcine kidney cell was the first choice to isolate and multiply PRCV AR310. The PRCV S1 gene of interest was amplified by RT-PCR and subsequently cloned into pMD18-T vector. The cloned S1 cDNA fragments were sequenced and homological analysis was done by DNAStar software. In S1 gene, the AR310 strain shared 100% nucleotide homology with PRCV DQ811787, 95.2% with M94096 and 95% with M94097 respectively. The amino acid homology of the tested PRCV S1 gene is 100% with DQ811787, 95.2% with M94096 and 95.6% with M94097. The amino acid among these strains were strictly conservative.
Keywords:PRCV
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